In-depth Understanding of a Nano-bio Interface Between Lysozyme and Au NP-immobilized N-doped Reduced Graphene Oxide 2-D Scaffolds

Overview

Journal Nanoscale Adv

Publisher Royal Society of Chemistry

Specialty Biotechnology

Date 2022 Sep 22

PMID 36132509

Authors

Karan Chaudhary

Krishan Kumar

Pannuru Venkatesu

Dhanraj T Masram

Affiliations

Soon will be listed here.

Abstract

In the present work, nitrogen-doped reduced graphene oxide (NrGO) was synthesized a hydrothermal treatment of graphene oxide (GO) in the presence of urea. Gold nanoparticles (Au(0) NPs) were immobilized over the surface of NrGO (Au(0)-NrGO). Characterization of the Au(0)-NrGO nanocomposite FT-IR spectroscopy, Raman spectroscopy, elemental mapping and XPS revealed the doping of N atoms during the reduction of GO. XRD and XPS studies confirmed the presence of Au(0) NPs and EDS analysis showed a 4.51 wt% loading of Au NPs in the Au(0)-NrGO nanocomposite. The morphology of Au(0)-NrGO was explored by SEM and TEM, which showed the presence of spherical Au metal NPs uniformly immobilized on the surface of NrGO. Further, studies on lysozyme (Lys) in the presence of Au(0)-NrGO by UV-visible, fluorescence, and circular dichroism spectroscopy revealed a conformational change in Lys and electrostatic interaction between Lys and Au(0)-NrGO. The DLS result showed an enhancement in the size of the Au(0)-NrGO and Lys conjugates. The Au(0)-NrGO-induced conformational changes in the structure of Lys resulted in a significant decrease in its activity at a certain concentration of Au(0)-NrGO. Moreover, the results showed that Lys favorably binds with the surface of Au(0)-NrGO, resulting in the formation of 2-D scaffolds possibly due to electrostatic and hydrophobic interactions, H-bonding, and interactions between the AuNPs and sulfur-containing amino acid residues of Lys. SEM exhibited the formation of conjugates in the form of 2-D scaffolds due to the biomolecular interactions between Lys and Au(0)-NrGO. The TEM studies revealed that Lys agglomerated around the Au(0) NPs immobilized on the surface of NrGO, which suggests the formation of a protein corona (PC) around the AuNPs. Furthermore, the favorable Au(0) NP-sulphur (PC) interaction was confirmed by the disappearance of the S-S stretching band in the Raman spectra. Overall, the results obtained provide insight into the nano-bio interface and formation of Au(0) NP-PC, which can be used for bioinspired applications, such as biosensing and imaging and the development of advanced functional Au NPs.

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References

Kanwa N, Patnaik A, De S, Ahamed M, Chakraborty A . Effect of Surface Ligand and Temperature on Lipid Vesicle-Gold Nanoparticle Interaction: A Spectroscopic Investigation. Langmuir. 2019; 35(4):1008-1020. DOI: 10.1021/acs.langmuir.8b03673. View

Wu T, Jiang Q, Wu D, Hu Y, Chen S, Ding T . What is new in lysozyme research and its application in food industry? A review. Food Chem. 2018; 274:698-709. DOI: 10.1016/j.foodchem.2018.09.017. View

Mogha N, Sahu V, Sharma R, Masram D . Reduced graphene oxide nanoribbon immobilized gold nanoparticle based electrochemical DNA biosensor for the detection of Mycobacterium tuberculosis. J Mater Chem B. 2020; 6(31):5181-5187. DOI: 10.1039/c8tb01604f. View

Lee D, Khatun Z, Lee J, Lee Y, In I . Blood compatible graphene/heparin conjugate through noncovalent chemistry. Biomacromolecules. 2011; 12(2):336-41. DOI: 10.1021/bm101031a. View

Zhang Y, Zhang J, Huang X, Zhou X, Wu H, Guo S . Assembly of graphene oxide-enzyme conjugates through hydrophobic interaction. Small. 2011; 8(1):154-9. DOI: 10.1002/smll.201101695. View

Shang W, Nuffer J, Muniz-Papandrea V, Colon W, Siegel R, Dordick J . Cytochrome C on silica nanoparticles: influence of nanoparticle size on protein structure, stability, and activity. Small. 2009; 5(4):470-6. DOI: 10.1002/smll.200800995. View

Lv M, Zhu E, Su Y, Li Q, Li W, Zhao Y . Trypsin-gold nanoparticle conjugates: binding, enzymatic activity, and stability. Prep Biochem Biotechnol. 2009; 39(4):429-38. DOI: 10.1080/10826060903210024. View

Strynadka N, James M . Lysozyme revisited: crystallographic evidence for distortion of an N-acetylmuramic acid residue bound in site D. J Mol Biol. 1991; 220(2):401-24. DOI: 10.1016/0022-2836(91)90021-w. View

Chakraborti S, Chatterjee T, Joshi P, Poddar A, Bhattacharyya B, Singh S . Structure and activity of lysozyme on binding to ZnO nanoparticles. Langmuir. 2009; 26(5):3506-13. DOI: 10.1021/la903118c. View

10.

Orecchioni M, Cabizza R, Bianco A, Delogu L . Graphene as cancer theranostic tool: progress and future challenges. Theranostics. 2015; 5(7):710-23. PMC: 4402495. DOI: 10.7150/thno.11387. View

11.

Vertegel A, Siegel R, Dordick J . Silica nanoparticle size influences the structure and enzymatic activity of adsorbed lysozyme. Langmuir. 2004; 20(16):6800-7. DOI: 10.1021/la0497200. View

12.

Liu J, Peng Q . Protein-gold nanoparticle interactions and their possible impact on biomedical applications. Acta Biomater. 2017; 55:13-27. DOI: 10.1016/j.actbio.2017.03.055. View

13.

Chung C, Kim Y, Shin D, Ryoo S, Hong B, Min D . Biomedical applications of graphene and graphene oxide. Acc Chem Res. 2013; 46(10):2211-24. DOI: 10.1021/ar300159f. View

14.

Lei C, Shin Y, Liu J, Ackerman E . Entrapping enzyme in a functionalized nanoporous support. J Am Chem Soc. 2002; 124(38):11242-3. DOI: 10.1021/ja026855o. View

15.

Zhang J, Zhang F, Yang H, Huang X, Liu H, Zhang J . Graphene oxide as a matrix for enzyme immobilization. Langmuir. 2010; 26(9):6083-5. DOI: 10.1021/la904014z. View

16.

Kuroki R, Weaver L, Matthews B . Structural basis of the conversion of T4 lysozyme into a transglycosidase by reengineering the active site. Proc Natl Acad Sci U S A. 1999; 96(16):8949-54. PMC: 17713. DOI: 10.1073/pnas.96.16.8949. View

17.

You C, Agasti S, De M, Knapp M, Rotello V . Modulation of the catalytic behavior of alpha-chymotrypsin at monolayer-protected nanoparticle surfaces. J Am Chem Soc. 2006; 128(45):14612-8. DOI: 10.1021/ja064433z. View

18.

Cha C, Shin S, Annabi N, Dokmeci M, Khademhosseini A . Carbon-based nanomaterials: multifunctional materials for biomedical engineering. ACS Nano. 2013; 7(4):2891-7. PMC: 3648999. DOI: 10.1021/nn401196a. View

19.

Bisht M, Kumar A, Venkatesu P . Refolding effects of partially immiscible ammonium-based ionic liquids on the urea-induced unfolded lysozyme structure. Phys Chem Chem Phys. 2016; 18(18):12419-22. DOI: 10.1039/c6cp01022a. View

20.

Aghili Z, Taheri S, Zeinabad H, Pishkar L, Saboury A, Rahimi A . Investigating the Interaction of Fe Nanoparticles with Lysozyme by Biophysical and Molecular Docking Studies. PLoS One. 2016; 11(10):e0164878. PMC: 5077090. DOI: 10.1371/journal.pone.0164878. View